A rapid protocol for the authentication of isolated differential display RT-PCR CDNAs

G Miele, R Slee, J Manson, M Clinton

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

The most time-consuming and problematic step in the overall DDRT-PCR technique is the confirmation that the isolated cDNA clone represents a differentially expressed gene. We have previously suggested that the majority of apparent false positives generated by DDRT-PCR do in fact result from the PCR reamplification of cDNA species which co-migrate with the cDNA of interest, and we have outlined a procedure to effectively eliminate these from further study. However, in situations where RNA is limiting, it is still desirable to confirm that a purified cDNA amplicon does, in fact, represent the originally observed differentially expressed gene prior to embarking on expression studies.
Original languageEnglish
Pages (from-to)245-55
Number of pages11
JournalPreparative Biochemistry & Biotechnology
Volume29
Issue number3
DOIs
Publication statusPublished - 1999

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